Annals. Food Science and Technology
2014
BIOCHEMICAL CHARACTERIZATION AND NUTRITIONAL PROPERTIES OF
Zizyphus lotus L. FRUITS IN AURES REGION, NORTHEASTERN OF ALGERIA
Mohamed Abdeddaim, Ourida Lombarkia, Ali Bacha, Djamel Fahloul, Djoumana Abdeddaim,
Radhia Farhat, Mouna Saadoudi, Yassine Noui and Adel Lekbir
Food Science Laboratory (LSA), Department of Food Technology, Veterinary and Agriculture Institute
University Hadj Lakhdar Batna, Algeria
Email: [email protected]
Abstract
The small fruits of Zizyphus lotus L., found in Aures region, northeastern of Algeria have been studied. The study was
realized on the pulp (P) and almond (A) separately. Levels of some nutrients in these parts were determined using
standard analytical methods. Crude protein, crude fat, crude fiber, ash, carbohydrate, pectin, moisture contents and
calorific values were in the (P) 3.80%, 1.32%, 8.41%, 3.28%, 65.90%, 3.78%, 12.27% and 16.341 Kj/g. In the (A) they
were 24.22%, 31.73%, 16.57%, 3.12%, 11.10%, 2.35%, 9.57% and 17.271Kj/g respectively. The (P) has a hard
consistency, with a sweet taste and very specific flavor. The TLC showed the presence of glucose, fructose and sucrose
like sugars. The profile of amino acids revealed the presence of cystein, glycin, arginin, serin, leucin, histidin and
alanin. Moreover, the almond is richer in fats. The fatty acids analysis by GC-MS revealed the presence of 15 fatty
acids; the unsaturated ones were at 71% with 49% of oleic acid of the total mixture, followed by 22% of linoleic acid.
The elemental analysis of the two parts of the fruit in mg/100g dry matter (DM) indicated that the (P) contained
appreciable levels of zinc (0.44), potassium (134.99), sodium (11.45), phosphorus (10.62), manganese (2.17),
magnesium (397.91) and iron (1.33). The (A) appears to be richer in these elements, mainly in phosphorus (24.00) and
magnesium (1349.06). These results revealed that this fruit presents a high nutritional value. Therefore, it should be
included in diets to satisfy needs of the body.
Keywords: Zizyphus lotus L., Jujube, Small fruit, Nutritional properties, Arid zone.
Submitted: 19.01.2014
Reviewed: 05.03.2014
lotus L (Ghedira et al., 1995; Renault et al.,
1997). These roots are used for treatment of
lung diseases, rheumatisms, arthritis, febrile
state and for healing in a folk medicine. Leaves
have a hypoglycemic effect and an antiseptic
activity (Epfraim et al., 1998; Abdel-Zaher et
al., 2005). The flowers infusion is used as
febrifuge and disinfectant for eyes (Sudhersan
and Hussain, 2003). Several effects of the fruits
of zizyphus have been reported like an antiage and anti-tumoral effects (Perdue and
Hartwell, 1976; Oh et al., 2004), the antidiarrheic and antiulcerogenic (Adzu et al.,
2002; Borgi et al., 2007), the antibacterial
effects (Ali et al., 2001; Nazif, 2002, Lahlou et
al., 2002). In the Aures region this fruit is
commonly used in food like a crudity,
especially by children. The aim of this study
was to investigate the nutritional quality of
these fruits.
1. INTRODUCTION
In the arid zone, the fruits of Zizyphus lotus L
are consumed by the autochthon population
and it constitutes a considerable food resource
for humans and animals in the food shortage
periods. The Zizyphus lotus L. is an
(Rhamnaceae) which grows in arid and
semiarid zone in the north of Sahara, its
endemic specie of the Aures region,
northeastern of Algeria. The plant called Sedra,
yields small fruits, the jujubes, they are named
locally N’Beg. Several parts of the plants of
Ziziyphus species are widely used in traditional
medicine for the treatment of several diseases
like gastrointestinal disturbance, liver troubles,
urinary infections, skin infections, insomnia,
diarrhea, and diabetes (Renault et al., 1997;
Glombitza et al., 1994; Le Croueour et al.,
2002). A cyclopeptidic alkaloids and saponins
are responsible for an antifungal and
antibacterial activities of the roots extract of Z
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Volume
15, Issue
1, 2014
Annals. Food Science and Technology
2014
Pulp
Fruit
Almond
Kernel
Figure 1. Fruit compartments of Z.lotus L
2. MATERIALS AND METHODS
The atomic absorption spectrometer Pye
Unicam sp 2900 with air acetylene Flame, was
used for the determination of the following
minerals; iron, magnesium, manganese and
zinc. The automatic extractor, Soxtec System
HT 1043 permited the extraction of fats from
the sample. A gas chromatograph GC-MS
Hewlett Packard HP 6890, coupled with a mass
spectrometer detector Hewlett Packard MSD
5973 with electronic impact was used for the
separation and identification of fatty acid
methyl esters.
2.1.Plant material
The fruits of Z. lotus L were collected from El
Mader region (Aures) during September 2009,
where this plant grows spontaneously. It was
kindly identified in the Agriculture Institute of
Batna. Edible parts, seeds and almonds were
manually isolated (Fig.1).
2.2. Physical characterization
Fifty fruits were aleatory chosen, the weight of
each fruit and its parts have been realized,
pulp, kernel and almond were weighed with a
precision scale Sartorius 1702. The measures
were determined with a digital caliper. The dry
mater was obtained by dehydration in oven
Memmert SLE 400 at 103±2 °C (Audigie et al.,
1984). The total energy has been realized with
an adiabatic calorimeter IkaWerk 5003 on the
dried powder. The refractive index of oil has
been determined with an Abee refractometer
Schmidt-Haensch.
2.3.1. Protein content
Total proteins were determined according to
the standard method of Kjeldahl (AFNORDGCCRF, 1995). The protein content is
obtained by the multiplication of total
nitrogen by 6.25 (Deymie et al.,1981).
2.3.2. Amino-acids identification
The identification of amino acids was realized
by thin layer chromatography (TLC) on silica
gel 0.25mm (Merck, Darmstadt, Germany).
The used mobile phase was butanol : acetic
acid: water (4 /1 /4). The sample was the
hydrolysat of protein in HCl 6N according to
the method reported by Audigie et al. (1984).
The revelation is realized by pulverization of
plate with 3% of Ninhydrine in acetone. It
2.3. Chemical characterization
A visible spectrophotometer, Shimadzu UV
120-01 was used to analyze the sample for
soluble sugars, and phosphorus. The extraction
apparatus Fibertec System Tecator 1010 has
been used to evaluate total fibers. A flame
spectrophotometer Jenwey PEP7 permited the
quantification of natrium and kalium.
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Figure 2. GC-MS Chromatogram of fatty acid methyl esters of almond oil of Z.lotus L fruit.
9
7
9
(1) C10: 0 – (2) C14: 0 – (3) C15: 0 – (4) C16: 0 – (5) C16: 1 – (6) C16: 1 – (7) C17: 0 – (8) C18: 0 – (9) C18: 1 – (10) C18: 2
9,12,15
11
(11) C18:3
– (12) C20: 0 – (13) C20: 1 – (15) C22: 0
shows the amino acids in blue purple spots on
the plate. The identification of amino acids is
based on the comparison of front reference Rf
of the amino-acids references (Merck,
Darmstadt, Germany) with those of samples.
esters
were
analysed
by
GC-MS,
chromatograph under the following conditions:
Polar capillary column Stabilwax (60 m, 0.25
mm, 0.25 µm). The column oven temperature
was programmed from 200 to 220 °C at 2
°C/minute and maintained for 30 mn. The
quantity of injected sample was 1µl. Helium
was the carrier gas with 0.5 ml/mn as flow. The
injection was realized in split less mode; the
injector temperature 250 °C, the transfer line
and the bloc were respectively at 280 °C and
230 °C. The mass spectrum was obtained in
ionization mode with electronic impact at 70
eV and scanning field from 50 to 600 uma at
2.83 scan/s. The recorded chromatogram is
represented in figure 2 and the results which
resume the fatty acids composition of the
sample were presented in table 3.
2.3.3. Extraction of lipids
About 50g of dried and powdered pulp or
almonds were extracted with 150ml of hexane
(Biochem Chemopharma, Georgia, USA) using
an automatic extractor (Soxtec System HT
1043) during 90 mn. The extract was collected
and the solvent evaporated in the water bath at
75 °C, the evaporation residue was dried in
oven up to constant weight. The saponification
index is determined in the usual way with
ethanolic KOH. An aliquot of 0.4g of oil is
dissolved in 20 ml of ethanolic KOH and
boiled under reflux during 45 mn. The excess
of potash was titrated with HCL at 0.2 N. The
refractive index of oil was obtained using the
Abee refratometer.
2.3.5. Free sugars analysis
The soluble sugars were obtained by
maceration of 2g of powder of edible part or
almonds in 50 ml of distilled water during one
hour with stirring. A 0.5g of CaCO3 were
added, the mixture was heated up to boiling.
After cooling, 0.5g of lead acetate was added
to the filtrate. After filtration, the extract was
2.3.4. Fatty acids identification
The fatty acid methyl esters were obtained by
the dissolution of 0.5g of fatty extract in 0.2 ml
of methanolic KOH 2N. The obtained methylic
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2014
used for the identification of soluble sugars by
TLC on silica gel 0.25mm using
Methylethylcetone / acetic Acid / Methanol
(3/1/1) as mobile phase. The revelation was
realized by spraying the plate with a mixture of
α-naphtol (Merck, Darmstadt, Germany) 0.25
g, absolute ethanol 50ml and 50ml sulfuric acid
20% and heating for 5 min. The comparison of
front reference of samples and the references
(Fluka Chemie GmbH, Germany) permits the
identification of soluble sugars (Multon, 1991).
The total fibers were determined with an
automatic apparatus by exhaustion using in
first a solution of sulphuric acid (Cheminova
International SA, Spain) 0.26 N during 30mn,
after cleaning with water a second exhaustion
with NaOH solution (Cheminova International
SA, Spain) 0.23N was realized. The difference
between the dry weight of the extract and the
weight after incineration gave the weight of
total fibers in the sample. The pectin was
determined by extraction on 10g of powdered
sample by 500ml of chlorhydric acid 0.1 N and
heating at 90 °C for one hour with a continuous
stirring. Then, the solution was left to settle for
12 hours and 5g of aluminium sulfate were
added to precipitate the pectin. The pH was
adjusted to 4 with ammonium hydroxide 1N.
The precipitate was filtered, dried and
weighted, results were summarized in table 2.
conditions. The results were summarized in
table 4.
2.3.7. Statistical analysis
Results regarding physical characteristics,
proximate analysis and elemental composition
were represented as means ± standard
deviation.
3. RESULTS AND DISCUSSIONS
The physical characteristics (figure 1) show
that the pulp constitutes 50% of fruit weight
and the almond 14% of seed weight (Table1).
Table 1: Physical characteristics of Z.lotus L
fruits.
Parameters
Values*
Fruit length (cm)
1.19±0.04
Fruit section (cm)
1.26±0.18
Pulp thickness(cm)
0.22±0.01
Kernel length(cm)
0.93±0.021
Kernel section(cm)
0.79±0.015
Pulp crud energy kJ/100g DM
1727.17±32
Almond crud energy kJ/100g DM 1625.73±28
Pulp proportion /Fruit (%)
50.36 ± 1.50
Almond proportion /Kernel (%)
14.58 ± 1.95
* Means± SD of five determinations.
The dry mater in level of 87% shows that the
edible part has a hard consistency and can be
stored for a long time. Measurements showed
that the jujube of Z.lotus L is a small fruit with
spherical form and brown color at maturity.
The pulp can yield 17.271kJ/g, a very
interesting quantity in regard to conventional
fruits. The refraction index showed that the oil
of Z. lotus L. is rich in oleic acid which was
confirmed by the GC MS analysis. The total
protein content is essentially present in seeds
with 14.22 % and shows a small quantity in the
pulp with 1.18% (Table 2). The amino acids
identified with TLC were serine, glycine,
alanin, leucin, histidin, arginin and cystein. The
lipids in the Z. lotus L fruit are localized in
seeds with a content of 29.73% of dry weight,
when a small amount is detected in the pulp
with 0.79%. The estimated unsaponifiable
fraction is 3.02% of dry weight.
2.3.6. Mineral analysis
The ashes were obtained by incineration of 1g
of
samples
in
muffle
furnace
Heraeus (MR.170) at 550 °C (Pinta et al.,
1980). The obtained ash was slowly moistened
with 3 ml of bidistilled water and 3 ml of
concentrated fluorhydric acid (Biochem
Chemopharma Georgia, USA). The filtrate was
adjusted to 100ml with bi-distilled water. The
analysis of minerals was realized by a visible
spectrophotometer (Shimadzu UV 120-01) for
the phosphorus, a flame photometer for
natrium and kalium and with atomic absorption
spectrometer Pye Unicam sp 2900 for
magnesium, manganese, iron and zinc using an
air acetylene flame atomization and a single
element lamps. For each element, a standard
curve is realized in the same analysis
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Table 2. Proximate analysis of Z.lotus L fruits.
almond. Their identification by TLC shows
that they are a mixture of glucose, fructose and
saccharose. The total amount was 16.57 % in
the edible part, probably is responsible for its
hardness. In the almond, fibers reached 16.57
% and are the important constituents of the
cuticles which cover the almond. The pectin is
present with 2.07 % in the edible part and
1.35% in the almond; it’s a poor fruit in pectic
compounds. The mineral matter present in the
edible part is 3.20 %, characterized by its
richness in phosphorus, natrium, kalium and
magnesium (Table 4).
Parameters
*Pulp
*Almond
Dry mater (%)
87.73 ± 0.20 92.43 ± 0.30
Ash%
3.20±0.15
92.43 ± 0.30
Crud Fats (%)
0.79 ± 0.02
29.73 ± 0.24
Soluble sugars (%) 10.55 ± 0.26 4.10 ± 0.23
Total fibres (%)
4.84 ± 0.55
16.57 ± 0.19
Pectins (%)
2.07 ± 0.33
1.35 ± 0.26
Crud protein (%)
1.18 ± 0.36
14.22 ± 0.89
* Means± SD of three determinations.
The fatty acids methyl esters analysis, by GCMS revealed the presence of 15 fatty acids.
Unsaturated fatty acids represent the major
components with 79.78 %. These unsaturated
fatty acids give the oil of Z. lotus L seeds a
high nutritive value, while the saturated ones
represent 20.22 % (Table 3). The major fatty
acids in this oil are in decreasing value, oleic
acid with 49.88 %, linoleic acid 22.97 %,
palmitic acid 9.05 %, stearic acid 7.10,
gadoleic acid 6.32 and arachidic acid 2.36 %.
Other fatty acids characterize this oil, like the
behenic acid. The soluble sugars represent
10.15 % in the edible part and 4.10% in the
Table 4. Elemental analysis of Z.lotus L fruit.
Minerals
mg/100g DM
*Pulp
*Almond
10.62±1.70
11.45±1.20
134.99±6.70
2.17±0.09
397.91±18.81
1.33±0.04
0.44±0.02
Phosphorus
Natrium
Kalium
Manganese
Magnesium
Iron
Zinc
24.00±0.63
17.41±1.63
97.92±7.86
7.84±0.93
1349.06±66.94
1.21±0.07
1.38±0.05
* Means± SD of three determinations.
Table 3. Fatty acids composition of almond oil of Z.lotus L fruit.
Retention
Time (mn)
Chemical denomination
Proportion % Mass
Usual nouns
5.581
Decanoic
Caprylic
C10
0.014
186
8.919
Tetradecanoic
Myristic
C14
0.084
242
10.401
Pentadecanoic
Pentadecylic
C15
0.024
256
12.411
Hexadecanoic
Palmitic
C16
12.874
7-Hexadecenoic
Hypogenic
9.025
270
7
0.058
268
9
0.134
268
0.077
284
7.106
298
49.882
296
22.973
294
0.409
292
2.367
326
C16 : 1
13.010
9-Hexadecenoic
Palmitoleic
C16 : 1
14.753
Heptadecanoic
Margaric
C17
18.150
Octadecanoic
Stearic
C18
19.306
21.233
9-Octadecenoic
9-12-Octadecadienoic
Oleic
Linoleic
C18 : 1
9
C18 : 2
9,12
9, 12,15
23.984
9-12-15-Octadecatrienoic
Linolenic
C18 : 3
27.624
Eicosanoic
Arachidonic
C20
11
29.261
cis11-Eicosenoic
Gadoleic
C20 : 1
6.328
324
34.615
Heneicosanoic
/
C21
0.047
340
44.225
Docosanoic
Behenic
C22
1.409
354
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The amount of phosphorus is more important
in the almond than in the edible part with 24.00
mg/100g DM, the same conclusion was found
by Largois (1994). This value is higher than
that found by Murdock (2002) with 65 mg/kg
of DM. Other varieties like Z.mauritiana and
Z. jujuba show a respective content of
phosphorus as 89.3 and 105mg/100g of DM
(Li et al., 2007). The magnesium content of
Z.lotus L as 397.91mg/100g of DM is more
important than that found by Murdock (2002).
The jujubes of Z.lotus L content of natrium is
11.45 mg/100g of DW. The presence of this
important quantity of natrium can be explained
by the presence of this plant in the dry and
salty soils in arid and semiarid zones. Kalium
in edible part of Z.lotus L fruit is in amount of
134.99mg/100g DW. This value is small with
regard to those found by Murdock (2002) for
the fresh jujube, of Z.mauritiana 250
mg/100g.The fruit of Z.lotus L can yield an
appreciable amount of oligoelements, iron,
manganese and zinc. The iron concentration in
the edible part of Z.jujuba (Zhao et al., 2006;
Zhao et al., 2007) is important with 3mg/100g
DW.The manganese content is higher than that
found by Murdock (2002) as 1mg/100g of DW
but it is in concordance with values reported by
Pinta et al.(1980). The zinc content of Z.lotus L
fruit is lower than that found by Murdock
(2002) in the fresh jujube. The proximate
biochemical composition in the Chinese jujube
was reported by Wei et al. (2007).
permits to classify this fruit as a functional
food. The essential oil constitutes a very
characteristic flavor of this fruit where it can be
used in food or in cosmetics. The study of fatty
fraction indicates that the oil in the fruit is
located in almond and has standard
characteristics of vegetable oils. The
biochemical characterization shows that this oil
is distinct by its richness in essential fatty
acids, thus it can be used as a food, in
pharmacology or in cosmetics. The edible part
cannot be considered as a source of protein
because its low content in this fraction but the
almond can be considered as a proteinous seed.
The studied proteins can compensate in part the
foodstuffs deficit shown in arid regions. The
studied fruit represents an appreciable source
of minerals, very appreciated and consumed by
children of local populations as crudeness in
the shortage period. This unconventional and
underutilized fruit has an interesting
biochemical composition which can be
valorized. This work can help to preserve the
specie by its integration as an agro-resource in
the sustainable development in arid regions of
the Sahara.
5. ACKNOWLEDGMENTS
The authors are grateful to Professor B.
Oudjehih, from Agriculture Institute of Batna
for the identification of specie, and to Drs Y.
Badja and R. Tir, from Haouari Boumedienne
University for the GC-MS analysis.
4. CONCLUSIONS
The study shows that the Z.lotus L. fruits are
dry product and the edible part is interesting
representing 50 % of fruit weight. The
proportion of almond in kernel is also
interesting, it could be valorized as a food or
like oleaginous. Because of its high energy
potential, this fruit can be considered as a high
energetic fruit. The moisture content is a good
parameter for its storage for a long time. The
study of pulp shows that the major compounds
of the biomass are sugars and they are also
important in the almond. The fiber content
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